Human Platelet Derived Growth Factor AB Recombinant

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Human Platelet Derived Growth Factor AB Recombinant

$70.00$3,500.00


accession P01127


Source Optimized DNA sequence encoding Human PDGF-AB mature chain was expressed in Escherichia Coli.
Molecular weight Recombinant Human PDGF-AB is a disulfide-linked heterodimer protein consisting of amino acid residue subunits(alpha+beta chains), and migrates as an approximately kDa protein under non-reducing conditions and reducing conditions in SDS-PAGE.
Purity >95%, as determined by SDS-PAGE and HPLC
Biological Activity The ED(50) was determined by the dose-dependentstimulation of thymidine uptake by BALB/cT3 was found to be in the range of1 ng/ml.

Protein Sequence MSIEEAVPAV CKTRTVIYEI PRSQVDPTSA NFLIWPPCVE VKRCTGCCNT SSVKCQPSRV HHRSVKVAKV EYVRKKPKLK EVQVRLEEHL ECACATTSLN PDYREEDTGR PRESGKKRKR KRLKPTSLGS LTIAE PAMIA ECKTR TEVFE ISRRL IDRTN ANFLV WPPCVEVQRC SGCCNNRNVQ CRPTQVQLRP VQVRKIEIVR KKPIFKKATV TLEDHLACKC ETVAAARPVT
Endotoxin Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation PDGF-AB was lyophilized from a.2 μm filtered solution in PBS pH.4.
Reconstitution A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers
Storage The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor P01730 CD4_HUMAN
Interactor P16234 PGFRA_HUMAN
Interactor P09619 PGFRB_HUMAN
Molecular function Developmental-protein
Molecular function Growth-factor
Molecular function Mitogen

Methods

Neural progenitor cell isolation and maintenance

  • All procedures were performed in sterile fashion in a class II biosafety cabinet.
  • A representative portion (2.5 cm) of three regions (thoracic, cervical and lumbar) of the spinal cord was used for Neural Progenitor Cell (NPC) isolation as described previously.
  • Briefly, tissue was diced and a single cell suspension was obtained by enzymatic dissociation of the tissue at 37 °C for approximately 30–40 minutes with 2.5 U/ml papain , 250 U/ml of DNase I , and 1 U/ml neutral protease .
  • After dissociation, the cell suspension was mixed with DMEM/F12 with 10% fetal bovine serum (FBS, , ), passed through a 70 μM filter, and centrifuged.
  • The cell pellet was resuspended in DMEM/F12 with 10% FBS and combined 1:1 with percoll (GE Healthcare, Piscataway, NJ).
  • The cell/percoll mixture was centrifuged at 20,000 g for 30 min at room temperature and the low buoyancy fraction (10 ml) above the red blood cell layer…
  • All procedures were performed in sterile fashion in a class II biosafety cabinet.
  • A representative portion (2.5 cm) of three regions (thoracic, cervical and lumbar) of the spinal cord was used for Neural Progenitor Cell (NPC) isolation as described previously.
  • Briefly, tissue was diced and a single cell suspension was obtained by enzymatic dissociation of the tissue at 37 °C for approximately 30–40 minutes with 2.5 U/ml papain , 250 U/ml of DNase I , and 1 U/ml neutral protease .
  • After dissociation, the cell suspension was mixed with DMEM/F12 with 10% fetal bovine serum (FBS, , ), passed through a 70 μM filter, and centrifuged.
  • The cell pellet was resuspended in DMEM/F12 with 10% FBS and combined 1:1 with percoll (GE Healthcare, Piscataway, NJ).
  • The cell/percoll mixture was centrifuged at 20,000 g for 30 min at room temperature and the low buoyancy fraction (10 ml) above the red blood cell layer was collected.
  • Cells were washed and resuspended in medium'>NPC medium containing DMEM/F12 supplemented with 10% FBS , 10% BIT9500 ( Cell , , ), 1% N2 supplement , 20 ng/ml of FGF-2 , 20 ng/ml of EGF , and 20 ng/ml of PDGF-AB .
  • NPCs were cultured on fibronectin coated plates and after 24 hours, the media was replaced with serum-free NPC medium.
  • Half of the medium was subsequently replaced every 2 days.
  • Cells were passed when 60–70% confluence was reached in about 3–4 weeks.

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